Certain cephalosporanic acid derivatives

ABSTRACT

3-(4-Oxo-2-thioxo-3-thiazolidinyl (lower) alkanamido)-1,4(cyclo-(1&#39;&#39;-carboxy)alkylenethio)azetidin-2-one derivitives possessing antibacterial activity are produced by the reaction of a 4-oxo-2-thioxo-4-thiazolidinyl-(lower)alkanoic acid derivative and a 6-aminopenicillanic or 7-aminocephalosporanic acid derivative in the presence of a condensing agent, or via an acid halide of the carboxylic acid.

United States Patent Teller et al.

[ Dec. 30, 1975 CERTAIN CEPHALOSPORANIC ACID DERIVATIVES Inventors: Daniel M. Teller, Devon; John H.

Sellstedt, Pottstown, both of Pa.

Assignee: American Home Products Corporation, New York, NY.

Filed: July 25, 1973 Appl. No.: 382,436

US. Cl 260/243 C; 260/239.1; 424/246;

424/271 Int. CL, C07D 501/22; C07D 501/34 Field of Search 260/243 C References Cited UNITED STATES PATENTS 11/1965 Flynn 260/243 C 3,268,523 8/1966 Raap et al 260/243 C Primary Examiner-V. D. Turner Attorney, Agent, or FirmRichard K. Jackson [57] ABSTRACT 3-[4-Oxo-2-thioxo-3-thiazolidiny1 (lower) alkanamido- ]-1,4-[cyclo-( l -carboxy)alkylenethio]azetidin-2-one derivitives possessing antibacterial activity are produced by the reaction of a 4-oxo-2-thioxo-4-thiazolidinyl-(lower)alkanoic acid derivative and a 6- aminopenicillanic or 7-aminocephalosporanic acid derivative in the presence of a condensing agent, or via an acid halide of the carboxylic acid 5 Claims, N0 Drawings 1 '-carboxy)alkylenethio]azetidin-2-ones CERTAIN CEPHALOSPORANIC ACID DERIVATIVES DESCRIPTION OF THE INVENTION In accordance with this invention, there is provided a group of azetidin-Z-one derivatives which are potent antibacterial agents generically termed 3-[4-oxo-2-thioxo-3-thiazolidinyl-(lower) alkanamido]-1,4-[cycloand the pharmaceutically acceptable salts thereof. These compounds present the structural formula:

' (cH cHcoNH wherein R is a member selected from the group consisting of H, alkyl of 1 to 6 carbon atoms, aryl of 6 to 10 carbon atoms, and aralkyl of 7 to 12 carbon atoms;

R is a member selected from the group consisting of -H and alkyl of 1 m6 carbon atoms;

R is a member selected from the group consisting of H, alkyl of l to 6 carbon atoms, monocyclic aryl of 6 to 10 carbon atoms, monocyclic aralkyl of 7 to 10 carbon atoms, OH, NH and CO H;

Y is a member selected from the group consisting of Y 4 H CH R or -CH C wherein R is H, (lower)alkanoyloxy,

-N -s-kJ CH3 and All R is a member selected from the group consisting of -H, an alkali metal cation and the ammonium ion; and

n is an integer from 0 to 5, inclusive.

The expression l,4-[cyclo-( l '-carboxy)alkylenethio], used in the generic name for the compounds of this invention, is intended to embrace the l-carboxy bridge member CO H as it appears in the preceding paragraph. The term (lower)alkyl is used to designate univalent aliphatic, hydrocarbon radicals containing from i to 6 carbon atoms, illustrative of which members are methyl, ethyl, n-propyl, i-propyl, n-butyl, t-butyl, n-pentyl, n-hexyl phoric, benzenesulfonic, toluenesulfonic, methylsulfonic, and ethylsulfonic acids, and the like.

The process of this invention, by which the above described compounds are prepared involves reacting the appropriately substituted precursor acid derivative of the formula:

N(CH2) CHCO-OH (01) with an appropriate 6-aminopenicillanic or 7- aminocephalosporanic acid derivative. The reaction procedes smoothly in an inert organic at a temperature of from about -40 to about +25C. When the free carboxylic acid is employed, the reaction is performed in the presence of a condensing agent such as carbonyl diimidazole; dicyclohexylcarbodiimide; dicyclohexylcarbodiimide in the presence of N-hydroxysuccimide or l-hydroxybenzotriazole; isobutylchloroformate; and

- the like. These and similar condensing agents which are operable in the preparation of the antibacterial agents of this invention are presented in Spencer et al., J Med. Chem. 9, pp 746-750 (1966); Micetich et al., J. Med. Chem. 15 pp. 333-335 (l972); Klausner et al., Synthesis, pp. 453-463 (1972) and U.S. Pat. No. 3,338,896.

Alternatively, the precursor carboxylic acid derivative may be converted by known means to an acid halide which is then used in aqueous medium to acylate the free amino group of either a tertiary amine salt or an alkali metal salt of the 7=aminocephalosporanic or -aminopenicillanic acid. In addition, the carboxylic acid halide precursor may be used to react in organic solution with either a tertiary amine salt or a silylated, phosphorylated or saccharinated derivatives of the 7-aminocephalosporanic acid or the -aminopenicillanic acid derivative.

The precursor carboxylic acid derivatives are prepared by the method of Zuber et al., Helv. Chim. Acta., 35, 1744 (1952); Minka, Farmat sevt. Zh. (Kiev) 18(5), 32-5 (1963); CA. 60, 5476a; or Brown, Chem. Rev., 61, 463 (1961). The generally. preferred scheme is:

KOH

the groups CH OC in which R is selected from the group consisting of H, halo, lower alkoxy and nitro, preferably R is hydrogen. In addition, the free amino group R may be protected by cyclization with the free carboxyl group with ethyl chloroformate and similar reagents to afford an acylating agent other than an acid halide.

The hydroxy protecting groups are tertiary butyl, trityl, benzyl, p-methoxybenzyl, and the like, the preferred blocking group being the benzyl radical. It is not essential that the hydroxyl group be protected during reaction with the 7-aminocephalosporanic acid or 6- aminopenicillanic acid derivatives, although it is preferred.

The a-carboxyl group may be protected as a tertiary butyl, benzyl, nitrobenzyl, methoxybenzyl, (e.g., pmethoxybenzyl, p-nitrobenzyl, 2,4-dimethoxybenzyl),

phenacyl, pht'halimidomethyl, B-methylthioethyl, diphenylmethyl, or 4-(methylthio)phenyl ester. Preferably the carboxyl group is protected in the form of a t-butyl ester, benzyl ester or substituted benzyl ester. Likewise, the free amino group and the free carboxy group in the azetidino reactant may be protected during the reaction with an alkali metal or tertiary amine, when operating in a mixed aqueous organic medium, or by monoor disilylation, phosphorylation or saccharination. In each case, the protective groups are readily removed by hydrolysis at the conclusion of the reaction.

The compounds of this invention are antibacterials effective against gram-positive and gram-negative test organisms as well as penicillin resistant staphylococcus at an inhibitory concentration at or below 250 micrograms per milliliter using the well-known and scientifically accepted agar serial dilution technique. The compounds have also been shown to be active in vivo. Thus,

the compounds of this invention are useful in the fields of comparative pharmacology and microbiology and may be used in the treatment of bovine mastitis, as growth promotors in animals and for the treatment of infections amenable to treatment with penicillins and cephalosporins.

The in vivo activity of the antibacterial agents of this invention was established by subcutaneously administering a single dose of the compound being tested to a randomized group of mice which had been infected six hours earlier with a specific infective agent, such as Escherichia coli, etc., via intraperitoneal injection of 0.5 milliliters of the infective agent in 5 per cent gastric mucin. The mice were then observed for 14 days and any deaths were recorded daily. The curative dose (CD was then reported in terms of the milligrams per mouse needed for complete control of the infective agent.

The following examples are presented for purposes of illustration and should not be construed as limitations upon the true scope of this invention. The biological activity data presented after each example illustrates the compounds activity against specific bacteria of the designated strain in terms of the minimum inhibitory concentration of the compound in micrograms per milliliter to completely inhibit the test organism. The abbreviations used in the examples are ACA- aminocephalosporanic acid; APA-aminopenicillanic acid; ADCA-amino-desacetoxycephalosporanic acid.

The abbreviations for the bacteria employed in the testing are as follows:

ST AU ll Staphylococcus aureus BA SU 6 -continued EXAMPLE 2 BO BR Bordetella hrochiseptica ES 1N Escherichia imermedia 6-[(4-Oxo-2-thioxo-3-thiazolidinyl)acetamid0]penicil- PR VU Proteus vulgaris l PR Ml Proteus mirablis 5 amc EN AE Emembac'e' aemgen Using the method described in Example 1 but substituting 6-APA (1.08 g, 0.005 moles) for 7-ACA and triturating the gummy product with diethyl ether gives EXAMPLE 1 the title compound, 0.50 g yellow solid, m.p. 7-[(4-Oxo-2-thioxo-3-thiazolidinyl)acetamido]ceph- 10 160 l70C. (decomp.); A 5.60 (shoulder), 5.71 alosporanic acid and 5.90 p, (shoulder); NMR has 1.46 and 1.60 ppm lets.

A solution of 4-oxo-2-thioxo-3-th1azo11d1ne acetic Smg acid'(0.96 g, 0.005 moles) in dry tetrahydrofuran 20 sgg g g z s for C3H5N3O5S3 ml) containing triet hylamine (0.70 ml) is cooled to 3 2 2 2 o Calc d: C, 40.91; H, 4.24; N, 10.40; S, 23.81.

10 C. Isobutyl chloroformate (0.66 ml) 18 added all at F C 40 H 4 N 10 S 20 98 once under nitrogen and the mixture stirred at 10C. under nitrogen another 10 minutes. A solution of 7-ACA (1.36 g, 0.005 moles) in tetrahydrofuran/wate'r Bacteria 3min MIC in #g/nfl (1:1, 18 ml) -conta1n1ng N,N-diisopropylethylamine BA SU 6633 976 (0.87 ml) is cooled to 0C and added rapidly to the ST AU I 1 above mixture. The temperature of the mixture is main- 5T AU SMITH tained at 5C. for 1 hour and then 20C. for another 52 hour. The tetrahydrofuran is evaporated in vacuo NE CA 8193 7.81 40C. A mixture of water (150 ml) and ethyl acetate 2 $2 x32 (40 ml) is added to the residue, the mixture Shaken PR VU 6896 31:3 thoroughly and the organic layer discarded. Ethyl ace- SP 9955 tate (200 ml) is added to the aqueous layer, the mixture cooled to 5C. and acidified to pH 2.5 with concentrated hydrochloric acid. The organic layer is sepa- EXAMPLE 3 rated, the aqueous layer extracted with ethyl acetate (60 ml), the vorgan: layer and extract combined, 7-[(4-Oxo-2-th1oxo-3 thiaaolidmyl)acetam1do]cephwashed with brine, dried over anhydrous sodium sulalosporamc Sodmm Salt fate and evaporated in vacuo 40C. The gummy To a slurry of 7-[(4-oxo-2-thioxo-3-thiazolidinylresidue is crystallized from acetone/benzene to give the 35 )acetamido]cephalosporanic acid (1.0 g, 0.0022 title compound, 0.88 g tan solid, mp. 300C; h moles) in water (20 ml) at 5C. is added 1.0 M aqueous 5.55, 5.70, 5.74, 5.83;.:.; NMR has 2.05 and 3.58 ppm sodium hydroxide (2.0 ml, 0.0020 moles) dropwise singlets. over 15 minutes. The solution is filtered and freeze- Elemental Analysis for C H N O S 0.2C H dried to give the title compound, 0.83 g orange solid, Calcd: C, 42.06: H, 3.53; N, 9.09; S, 20.82. 40 mp. 165180C (decomp.), A 5.68 and 6.21;/.; Found: C, 41.90; H, 3.62; N, 9.10; S, 19.72. NMR has 2.02 ppm singlet;

' Elemental Analysis for C H N O,S Na H O Calcd: C, 36.43; H, 3.46; N, 8.51; S, 19.45; H O, Bacteria Strain MIC in tglml y 547 BA SU 6633 5 Found: C, 36.88; H, 3.33; N, 8.55; S, 18.98; H O, ST AU 6538P .488 1 ST AU SMITH .488 ST AU CHP 1.95 ST AU 53-180 .976 EXAMPLE 4 it ig 1 3-Methy1-8-oxo-7-[2-(4-oxo-2-thioxo-3-thiazo1idinyl- 30 BR 4617 )acetamidol-S-thia-1-azabicyclo[4.2.0loct-2-ene-2- ES 1N 65-1 125 carboxylic acid. PR VU 6896 125 EN AE 13048 250 Using the method described in Example 1 but substi- 5 3: 3g; 2g 5 tuting 7-ADCA 1.07 g, 0.005 moles) for 7-ACA and ES c0 9637 2:5 triethylamine (0.70 ml.) for N,N-diisopropylethyla- 55 mine and slurrying the product with diethyl ether gives the title compound; 0.15 g tan solid, mp. 210-216C.; The in vivo data for the product of Example 1, in mu' 5-55, 5-72, l NMR has and terms of the curative dose (CD expressed as the dose PP singletsin milligrams per mouse via subcutaneous administra- Another g of P1001119t obtamed y flltermg Off i i as f llow 60 the solid formed after acidification to pH 25 with concentrated hydrochloric acid in the work up.

Bacteria Strain CD, mg/mouse ES CO 920 -l7 Bacteria Strain MlC in pig/ml KL PN KL-l 1.59 PR M1 3 1.36 BA SU 6633 3.90 PR VU 347 4.31 ST AU SMITH 7.81 ST AU SMITH 7.81 ST AU CHP 31.3 ST AU 53-180 15.6

-continued Bacteria Strain MIC in ,ug/ml SA PA 11737 125 KL PN 11737 125 KL PN 10031 250 PR VU 6898 250 EXAMPLE 7-[3-(4-Oxo-2-thioxo-3-thiazolidinyl)propionamido]- 10 cephalosporanic acid Using the method described in Example 1 but substituting 4-oxo-2-thioxo-3-thiazolidine propionic acid (1.03 g, 0.005 moles) for 4-oxo-2-thioxo-3-thiazolidine acetic acid and triturating the product with diethyl ether gives the title compound, 1.45 g off-white solid, m.p. 137l41C. (decomp.) )t 5.56, 5.75, 6.03 2; NMR has 2.03 ppm singlets Elemental Analysis for C 1-1 N O,S 1/4 CH C1-1 OC1-l CH Calcd: C, 42.72; H, 4.11; N, 8.79, S, 20.11 Found: C, 42.60; H, 4.24; N, 8.62; S, 18.36

Bacteria Strain MlC in ug/ml BA SU 6633 .061 ST AU 6538? .244 ST AU SMITH .122 ST AU CH? .976 ST AU CH? .976 5 ST AU 53-80 .976 NT CA 8193 31.3 ES CO 9637 15.6 ES IN 65-1 125 SA PA 11737 1.95 EN AE 13048 125 KL PN 10031 7.81 BO BR 4617 31.3 PR VV 6896 7.81

EXAMPLE 6 7-[3-Phenyl-2-(4-oxo-2-thioxo-3-thiazolidinyl)propionamido]cephalosporanic acid Using the method describied in Example 1 but substituting 3-phenyl-2-[4-oxo-2-thioxo-3-thiazolidinyl]propionic acid (1.68 g. 0.005 moles) for 4-oxo-2-thioxo-3- thiazolidine acetic acid gives the title compound as a gum, )t 5.59, (shoulder), 5.78 ;1.;NMR has 1.97 and 7.29 ppm singlets.

What is claimed is:

l. A compound of the formula:

N(C cHcoNH-F 4 s CH2R 5 C0 12 in which R is a member selected from the group consisting of H, alkyl of 1 to 6 carbon atoms, aryl of 6 to 10 carbon atoms and aralkyl of 7 to 12 carbon atoms; R is a member selected from the group consisting of H and alkyl of 1 to 6 carbon atoms;

R is a member selected from the group consisting of [2-(4-oxo-2-thioxo-3-thiazolidinyl)acetamido]-5-thial-azabicyclo-[4.2.0]oct-2-ene-2-carboxylic acid, or an alkali metal or the ammonium salt thereof.

5. A compound of claim 1 which is 7-[3-phenyl-2-(4- oxo-2-thioxo-3-thiazolidiny1)propionamido]cephalosporanic acid, or an alkali metal salt or the ammonium salt thereof. 

1. A COMPOUND OF THE FORMULA:
 2. A compound of claim 1 which is 7-((4-oxo-2-thioxo-3-thiazolidinyl)acetamido)cephalosporanic acid, or an alkali metal or the ammonium salt thereof.
 3. A compound of claim 1 which is 7-(3-(4-oxo-2-thioxo-3-thiazolidinyl)propionamido)cephalosporanic acid, or an alkali metal or the ammonium salt thereof.
 4. A compound of claim 1 which is 3-methyl-8-oxo-7-(2-(4-oxo-2-thioxo-3-thiazolidinyl)acetamido)-5-thia-1 -azabicyclo-(4.2.0)oct-2-ene-2-carboxylic acid, or an alkali metal or the ammonium salt thereof.
 5. A compound of claim 1 which is 7-(3-phenyl-2-(4-oxo-2-thioxo-3-thiazolidinyl)propionamido)cephalosporanic acid, or an alkali metal salT or the ammonium salt thereof. 